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Identification of P2Y12-dependent and -independent mechanisms of glycoprotein VI–mediated Rap1 activation in platelets by Mark K. Larson, Hong Chen, Mark L. Kahn, Anne M. Taylor, Jean-Etienne Fabre, Richard M. Mortensen, Pamela B. Conley, and Leslie V. Parise Blood Volume 101(4):1409-1415 February 15, 2003 ©2003 by American Society of Hematology CVX activates Rap1 in platelets in a concentration- and time-dependent manner.(A) Equal numbers of washed human platelets were stimulated with the indicated concentrations of CVX (left panel) or fibrillar type I collagen (right panel) for 1 minute. Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology CVX, but not collagen, treatment of RBL-2H3 cells stimulates Rap1 activation in an FcRγdependent manner.RBL-2H3 cells stably transfected with wild-type GPVI (GPVI.ori), a mutant GPVI that fails to associate with FcRγ (GPVI.R272L), or untransfected RBL-2H3 ... Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology Collagen, but not CVX, requires α2β1 to fully stimulate Rap1 in human platelets.Washed human platelets were treated with control buffer, 10 μg/mL control IgG1, or 10 μg/mL of the α2-blocking mAb P1E6 for 15 minutes and then stimulated with either fibrillar ... Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology Full CVX-mediated Rap1 activation is dependent on ADP but not TXA2 signaling and specifically requires the P2Y12 ADP receptor in human platelets.(A) Washed human platelets were pretreated with dimethyl sulfoxide (DMSO; control) or the indicated concentratio... Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology Differential effects on Rap1 activation in Gαi2 −/− murine platelets in response to ADP and CVX.(A) Washed wild-type (wt) and Gαi2-deficient murine platelets were stimulated with 50 μM ADP or 500 ng/mL CVX for 1 minute before lysis. Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology Effect of 2-MeSAMP on Rap1 activation in wild-type and Gαi2 −/− platelets stimulated with ADP and CVX.(A) Washed wild-type (wt) or Gαi2-deficient murine platelets were treated with 2MeSAMP or PBS for 15 minutes, followed by CVX (500 ng/mL, n = 4) or ADP (5... Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology Epinephrine augments CVX-mediated Rap1 activation in Gαi2-deficient murine platelets and 2MeSAMP–treated human platelets.(A) Washed Gαi2-deficient murine platelets were stimulated with either CVX or epinephrine (2 μM) or CVX and epinephrine for 1 minute. Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology PI3K is a part of the ADP-independent component of GPVI-mediated Rap1 activation.(A) Human platelets were pretreated with DMSO or BAPTA-am(BAP, 20 μM), PP2 (30 μM), or LY 294002 (LY, 100 μM), 10 minutes prior to a 1-minute stimulation with CVX (250 ng/mL) o... Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology CVX-induced platelet aggregation is mediated by ADP-dependent and -independent components.Aggregation traces from wild-type (wt, A-B) or Gαi2 −/− (C) murine platelets treated with either control buffer (A,C) or the ADP receptor antagonists 2-MeSAMP (50 μM) ... Mark K. Larson et al. Blood 2003;101:1409-1415 ©2003 by American Society of Hematology
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